Fig. 6

Calpastatin/Calpain proteolysis system influences cell migration velocity and F-actin behavior in Fam20c-deficient cells. A Gene expression of Calpastatin, Calpain 1, and Calpain 2 in OB Fam20c^KO and OB Fam20c^KO treated with CaCl₂. ^*P < 0.05, ^**P < 0.01, ^****P < 0.0001. B Western blot analysis of Calpastatin, Calpain 1, and Calpain 2 expression in OB Fam20c^KO and OB Fam20c^KO treated with CaCl₂. GAPDH served as an internal control. C Gene expression of Calpastatin, Calpain 1, and Calpain 2 in OB Fam20c^f/f, OB Fam20c^KO, and OB Fam20c^KO treated with different concentrations of CaCl₂. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, ^****P < 0.0001, NS: no significance. D Phos-Tag sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) (P-tag) demonstrates effects of CaCl₂ on Calpastatin phosphorylation levels in OB Fam20c^f/f, OB Fam20c^KO, and OB Fam20c^KO treated with CaCl₂. Tubulin served as an internal control. E Casein zymography profiles of Calpain 1 and Calpain 2 in OB Fam20c^f/f, OB Fam20c^KO, and OB Fam20c^KO treated with CaCl₂. F Wounding healing assays of OB Fam20c^f/f, OB Fam20c^KO, and OB Fam20c^KO treated with CaCl₂, images were acquired 0 h and 24 h after the scratch. Scale bar = 100 μm. G Gene expression of Rac1, RhoA, and Cdc42 in OB Fam20c^KO and OB Fam20c^KO treated with CaCl₂. ^*P < 0.05, ^**P < 0.01. H Western blot analysis of Cdc42 expression in OB Fam20c^f/f, OB Fam20c^KO and OB Fam20c^KO treated with CaCl₂. GAPDH served as an internal control